Optogenetic Fine-Tuning of <i>Sus scrofa</i> Basic Fibroblast Growth Factor Expression in <i>Escherichia coli</i>
Basic fibroblast growth factor (bFGF) is a crucial protein with diverse applications in biotechnology and medicine. This study aims to investigate the use of EL222-based optogenetic control systems to fine-tune the expression of porcine (<i>Sus scrofa</i>) bFGF in <i>Escherichia co...
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MDPI AG
2024-11-01
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| Series: | Fermentation |
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| author | Fanqiang Meng Zhimin Xu Xia Fan Zhisheng Wang Libang Zhou |
| author_facet | Fanqiang Meng Zhimin Xu Xia Fan Zhisheng Wang Libang Zhou |
| author_sort | Fanqiang Meng |
| collection | DOAJ |
| description | Basic fibroblast growth factor (bFGF) is a crucial protein with diverse applications in biotechnology and medicine. This study aims to investigate the use of EL222-based optogenetic control systems to fine-tune the expression of porcine (<i>Sus scrofa</i>) bFGF in <i>Escherichia coli</i>. The bioactivity and the productivity of blue light-induced bFGF were demonstrated to be comparable to those achieved using a conventional T7-expression system. Secondly, through systematic optimization of regulatory elements, optimal expression of bFGF was achieved using a medium-strength promoter for EL222 expression, a strong RBS upstream of the bFGF gene, and an optimized C120 configuration within the blue light-inducible promoter. Moreover, various parameters of blue light illumination during fermentation were investigated, including initial cell density, light intensity, illumination duration, and pulsed illumination patterns. The results identified optimal conditions for maximizing bFGF yield in <i>E. coli</i>, specifically an initial OD<sub>600</sub> of 0.6, 800 lux blue light intensity, and 8 h total illumination in a 2 h on/off pattern. Overall, this successful implementation of optogenetically controlled bFGF expression in <i>E. coli</i> serves as a proof-of-concept for light-responsive systems in industrial biotechnology, highlighting the potential of optogenetic control for biologically active protein production. |
| format | Article |
| id | doaj-art-c701c9aaa63e44388ad525426694b8df |
| institution | DOAJ |
| issn | 2311-5637 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Fermentation |
| spelling | doaj-art-c701c9aaa63e44388ad525426694b8df2025-08-20T02:53:30ZengMDPI AGFermentation2311-56372024-11-01101261210.3390/fermentation10120612Optogenetic Fine-Tuning of <i>Sus scrofa</i> Basic Fibroblast Growth Factor Expression in <i>Escherichia coli</i>Fanqiang Meng0Zhimin Xu1Xia Fan2Zhisheng Wang3Libang Zhou4College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, ChinaDepartment of Food and Drug Engineering, Shandong Vocational Animal Science and Veterinary College, Weifang 261061, ChinaCollege of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, ChinaNational Research Center of Engineering and Technology for Veterinary Biologicals, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, ChinaCollege of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, ChinaBasic fibroblast growth factor (bFGF) is a crucial protein with diverse applications in biotechnology and medicine. This study aims to investigate the use of EL222-based optogenetic control systems to fine-tune the expression of porcine (<i>Sus scrofa</i>) bFGF in <i>Escherichia coli</i>. The bioactivity and the productivity of blue light-induced bFGF were demonstrated to be comparable to those achieved using a conventional T7-expression system. Secondly, through systematic optimization of regulatory elements, optimal expression of bFGF was achieved using a medium-strength promoter for EL222 expression, a strong RBS upstream of the bFGF gene, and an optimized C120 configuration within the blue light-inducible promoter. Moreover, various parameters of blue light illumination during fermentation were investigated, including initial cell density, light intensity, illumination duration, and pulsed illumination patterns. The results identified optimal conditions for maximizing bFGF yield in <i>E. coli</i>, specifically an initial OD<sub>600</sub> of 0.6, 800 lux blue light intensity, and 8 h total illumination in a 2 h on/off pattern. Overall, this successful implementation of optogenetically controlled bFGF expression in <i>E. coli</i> serves as a proof-of-concept for light-responsive systems in industrial biotechnology, highlighting the potential of optogenetic control for biologically active protein production.https://www.mdpi.com/2311-5637/10/12/612optogeneticEL222bFGFblue light<i>Escherichia coli</i> |
| spellingShingle | Fanqiang Meng Zhimin Xu Xia Fan Zhisheng Wang Libang Zhou Optogenetic Fine-Tuning of <i>Sus scrofa</i> Basic Fibroblast Growth Factor Expression in <i>Escherichia coli</i> Fermentation optogenetic EL222 bFGF blue light <i>Escherichia coli</i> |
| title | Optogenetic Fine-Tuning of <i>Sus scrofa</i> Basic Fibroblast Growth Factor Expression in <i>Escherichia coli</i> |
| title_full | Optogenetic Fine-Tuning of <i>Sus scrofa</i> Basic Fibroblast Growth Factor Expression in <i>Escherichia coli</i> |
| title_fullStr | Optogenetic Fine-Tuning of <i>Sus scrofa</i> Basic Fibroblast Growth Factor Expression in <i>Escherichia coli</i> |
| title_full_unstemmed | Optogenetic Fine-Tuning of <i>Sus scrofa</i> Basic Fibroblast Growth Factor Expression in <i>Escherichia coli</i> |
| title_short | Optogenetic Fine-Tuning of <i>Sus scrofa</i> Basic Fibroblast Growth Factor Expression in <i>Escherichia coli</i> |
| title_sort | optogenetic fine tuning of i sus scrofa i basic fibroblast growth factor expression in i escherichia coli i |
| topic | optogenetic EL222 bFGF blue light <i>Escherichia coli</i> |
| url | https://www.mdpi.com/2311-5637/10/12/612 |
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